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2024 Update: Echinacea hybrids (exPt 6,7,9) and Echinacea pallida flowering phenology

Echinacea pallida is a species of Echinacea that is not native to Minnesota. It was mistakenly introduced to our study area during a restoration of Hegg Lake WMA around 2006 (Stuart’s recollection). This is concerning, because we don’t know how a similar species may impact or local Echinacea angustifolia! Will they hybridize? Could pallida outcompete angustifolia? Ever since pallida have started springing up, Team Echinacea has visited the pallida restoration, taken flowering phenology, and collected demography on the non-native plant. We have decapitated all flowering E. pallida each year to avoid cross-pollination with the local Echinacea angustifolia. Each year, we record the number of heads on each plant and the number of rosettes, collect precise GPS points for each individual, and cut off all the heads before they produce fruits.

Echinacea pallida flowering: 

This year, we cut E. pallida heads on June 26th. Overall, we found and shot 172 flowering E. pallida plants with 512 normal heads in total, averaging 2.98 heads per plant, though the max was 20 on a single plant! These non-native plants were hearty with an average rosette count of 7.20 rosettes and an astounding individual with a maximum of 88 rosettes. We did not take phenology data on E. pallida this year.

Team 2024 returns to their vehicles after flagging, taking demography data on, and decapitating Echinacea pallida at Hegg Lake WMA
  • Start year: 2011
  • Location: Hegg Lake WMA (MN DNR)
  • Overlaps with: 
  • Data collected: 
    • Demography data: head counts, rosette counts, etc.
      • data in aiisummer2024 repo: ~/aiisummer2024/demo/demoGood2024.txt
    • Spatial location for every flowering E. pallida
      • data in aiisummer2024 repo: ~/aiisummer2024/surv/survGood2024.txt
  • Samples collected:
    • Echinacea pallida were not collected: decapitated heads were left on the ground next to the plants
    • A flowering echinacea at the aptly named nearby remnant “near pal” looked suspiciously like a hybrid (more robust than an angustifolia). We put a pollinator exclusion bag on the single head to prevent pollen spread, and later harvested the head and brought it back to the lab, where it is currently in the seed dryer. Keep an eye on tag 29239 in the future
  • Products:
    • None… yet! Besides a prairie with significantly less E. pallida reproduction

You can find more information about E. pallida flowering phenology and previous flog posts on the background page for the experiment.

exPt06: 

Experimental plot 6 was the first E. angustifolia x E. pallida hybrid plot planted by Team Echinacea. A total of 66 Echinacea hybrids were originally planted. All individuals have E. angustifolia dams and E. pallida sires. In 2024, we visited 23 positions, 4 of which were can’t find year 3 in 2023 and didn’t get their final double check. We found living plants at all positions but those four (so, 19)! Last year, for the first time, 3 plants flowered in this plot. This year, no plants flowered.

  • Start year: 2011 (crossing) and 2012 (planting)
  • Location: Wagenius property
  • Overlaps with:
    • Common garden experiment
  • Data collected: 
    • Measure data (status, size, etc.)
      • data in SQL database
  • Samples collected:
    • None (no flowering plants)
  • Products:
    • None… yet!

You can find more information about experimental plot 6 and previous flog posts about it on the background page for the experiment.

exPt07: 

Experimental plot 7 is the second E. pallida x E. angustifolia plot. It contains conspecific crosses of each species as well as reciprocal hybrids, totaling 294 pdeigreed individuals. We took phenology records between July 10th and July 18th. There were 42 flowering plants this year; from these we harvested 87 heads. Heads in this plot were covered by pollinator exclusion bags during the growing season to prevent cross-pollination with nearby Echinacea populations.

Stuart demonstrates proper measuring technique in exPt07
  • Start year: 2012 (crossing) and 2013 (planting)
  • Location: Hegg Lake WMA (MN DNR)
  • Overlaps with: 
    • Common garden experiment
  • Data collected: 
    • Phenology data (dates of flowering stages)
      • data in cgData repo: ~/cgData/summer2024/exPt79Phenology
    • Measure data (status, size, etc.)
      • data in SQL database
    • Harvest data (IDs of harvested heads, missing achenes, etc)
      • detailed data in dropbox: dropbox/CGData/140_reconcile/reconcile2024/reconcileOut/2024harvestListReconciledExport.csv
      • data in echinaceaLab package (hh.2024)
  • Samples collected:
    • 87 heads harvested
      • at CBG for processing
  • Products:
    • None… yet!

You can find more information about experimental plot 7 and previous flog posts about it on the background page for the experiment.

exPt09: 

There were originally 745 seedlings planted in exPt09. Experimental plot 9 is a hybrid plot, but, unlike the other two hybrid plots, we do not have a perfect pedigree of the plants. That is because the E. angustifolia and E. pallida maternal plants used to generate seedlings for exPt09 were open-pollinated. At this point, some but not all plants in this plot were tested for paternity, revealing that there are some hybrids. This year, we took phenology records between July 9th and July 18th. During measuring, we searched at 292 positions and found evidence of 234 living plants in 2024. Of these individuals, 70 were flowering. We harvested 110 heads from this plot! Heads in this plot were covered by pollinator exclusion bags during the growing season to prevent cross-pollination with nearby Echinacea populations.

The team runs out reel tapes to aid in measuring exPt09
  • Start year: 2014
  • Location: Hegg Lake WMA (MN DNR)
  • Overlaps with: 
    • Common garden experiment
  • Data collected: 
    • Phenology data (dates of flowering stages)
      • data in cgData repo: ~/cgData/summer2024/exPt79Phenology
    • Measure data (status, size, etc.)
      • data in SQL database
    • Harvest data (IDs of harvested heads, missing achenes, etc)
      • detailed data in dropbox: dropbox/CGData/140_reconcile/reconcile2024/reconcileOut/2024harvestListReconciledExport.csv
      • data in echinaceaLab package (hh.2024)
  • Samples collected:
    • 110 heads harvested
      • at CBG for processing
  • Products:
    • None… yet!

You can find out more information about experimental plot 9 and flog posts mentioning the experiment on the background page for the experiment.

2024 Update: Ground-nesting bees in prairie remnants and restorations

During the summer of 2024, Team Echinacea completed the second year of its ENRTF funded project to better understand how prescribed fire influences ground nesting bee habitat, food resources, and diversity. Understanding the associations between land management methods and ground nesting bees is essential for providing reccomendations to policymakers and practitioners interested in native bee conservation.

We surveyed solitary bee diversity and nesting habitat before and after prescribed fires in a subset of 30 prairie remnants and 15 prairie restorations to determine how prescribed fire affects solitary bee nesting habitat and abundance. We used emergence traps to sample the community of solitary ground nesting bees. This was complemented by detailed measures of soil and litter to characterize how prescribed burning influences the nesting habitat (read more here).

2024 REU student Zach Zarling deploys an emergence trap at a site near Hoffman, Minnesota

We deployed emergence traps at our random “burn and bee points”(BBPTs) in prairie remnants and restorations from early June to mid September. Our deployments spanned three rotations (4-6) of BBPTs and we put out a total of ~1,159 emergence traps. On reccomendation from Dr. Alex Harmon-Threatt, we also performed 10 minute “pollard walks” on deployment to estimate the number of foraging bees at each site. These foraging numbers will be compared to nesting incidence as part of Ian Roberts’ thesis project.

As of December 21st, specimens caught in this year’s deployments have been pinned, labeled, and transported from Chicago Botanic Garden to the University of Minnesota, where Zach Portman, a bee taxonomist, will identify them. Team Echinacea also collected lots of non-bee bycatch while processing specimens collected in the traps: including millipedes, flies, and even a prairie skink! To avoid wasting these specimens, we plan to categorize this bycatch into broad taxonomic groups (like Dipterans, Orthopterans, etc) and examine potential associations between our experimental treatments and general arthropod diversity across our study sites.

Pinned specimen from 2024’s emergence trapping, likely an Agapostemon virescens.

While working on pinning and processing specimens, Ian Roberts produced a poster containing analyses from the 2023 emergence trapping data to present at Entomology 2024. The poster can be viewed here. Future data analyses will feature data from both sampling years, as well as microhabitat measurments and and diversity indices.

2024 Update: Microhabitat in prairie remnants and restorations

During summer 2024, Team Echinacea continued to collect data on local environmental conditions in order to understand which environmental factors are associated with good habitat for ground-nesting bees. These data complement emergence trapping for our ENRTF funded research on fire’s influence on ground nesting bees habitats. We sampled local environmental conditions near randomly placed “burn and bee points” (BBPTs) in prairie remnants and restorations.

A deployed emergence trap next to a set of marking flags. Microhabitat data was sampled within a meter of these marking flags.

Unlike the 2023 season, we did not collect data on light levels at BBPTs this year. Instead, we measured soil temperature just under the surface using a digital thermometer, along with soil compaction via a penetrometer and litter depth via a meter stick.

Team Echinacea conducted microhabitat assessments for remnant prairies at rotation 4 BBPTs. Over the summer, we took microhabitat assessment measurements at a total of 241 measurments.

  • Start year: 2024
  • Location: prairie remnants and restorations in Solem Township, MN
  • Overlaps with: ground nesting bees
  • Data collected: Surface soil temperature (°C), soil compaction (psi), and litter depth (cm)
  • Samples or specimens collected: NA
  • Products: Poster presented at Entomology 2024 in early November: link to post containing poster

ACE progress update: polOb 2024

Another batch we’re prioritizing in the lab is the pollinator observation (or polOb) experiment. The goal of this experiment is to learn more about the effects of fire on native bees and their behaviors. We spent time this summer observing Echinacea at 10 paired sites, half of which burned before the 2024 growing season. We then harvested heads from our observation plots for processing in the lab. Because we’re specifically interested in learning about pollination, heads in this group will be streamlined all the way through to the classifying steps to learn about seed set, and we are less concerned about counting. Here’s our progress so far:

We are roughly a third of the way through rechecking. We’ll be pausing work briefly on this batch to get the exPt01 2024 experiment rechecked and uploaded for counting ASAP

NOTE: Funding for this project was provided by the Minnesota Environment and Natural Resources Trust Fund as recommended by the Legislative-Citizen Commission on Minnesota Resources (LCCMR).

ACE progress update: exPt01 2024

We have a few experiments that we’re prioritizing here in the lab. We want to get the heads in these experiments through our ACE process as quickly as possible! One of these experiments is known by many names: q1, qGen, qGen1, big batch, etc. The experiment in question was planted in experimental plot 1 in 2003 and was designed to investigate the heritability of fitness using a quantitative genetics approach. We’re looking to redo the analysis we have on this experiment with several new years of data, including 2024!

This year, very few plants in exPt01 flowered, so all experiments in that plot were lumped into one for ACE processing. I booted up Alex’s ACE progress visualization machine to see what our progress on this batch looks like so far! We only need to get this batch through counting, so we can skip the randomizing/xray/classifying steps.

Finished with cleaning! Rechecking (here cleanQC) is up next.

2024 Update: Aphid addition and exclusion

The aphid addition and exclusion experiment was started in 2011 by Katherine Muller. The original experiment included 100 plants selected from exPt01 that were each assigned to have aphids either added or excluded across multiple years. The intention is to assess the impact of the specialist herbivore Aphis echinaceae on Echinacea fitness.

In 2024, 41 of the original 100 plants were alive, two of which flowered. However, we did not see any aphids anywhere while measuring exPt01. We have not conducted fieldwork for this experiment since 2022, when team members Emma Reineke and Kennedy Porter were in charge of the experiment and did not find any aphids in exPt01, so they introduced a new population of Aphid echinaeceae into the plot. Learn more in the 2022 summer aphid update.

Aphids Wyatt Mosiman saw in exPt02 this summer. Wrong plot, aphids!
  • Start year: 2011
  • Location: exPt01
  • Overlaps with: Phenology and fitness in P1
  • Data collected: 
    • Plant survival and measurements were recorded as part of our annual surveys in P1 and eventually will be found in our SQL database.
  • Samples collected:
    • 2 heads from plants included in this experiment are at the Chicago Botanic Garden awaiting processing: AD-1728 and AZ-1744 in the exPt01 2024 batch
  • Products:
    • Andy Hoyt’s poster presented at the Fall 2018 Research Symposium at Carleton College
    • 2016 paper by Katherine Muller and Stuart Wagenius on aphids and foliar herbivory damage on Echinacea
    • 2015 paper by Ruth Shaw and Stuart Wagenius on fitness and demographic consequences of aphid loads

You can read more about the aphid addition and exclusion experiment, as well as links to prior flog entries mentioning the experiment, on the background page for this experiment.

2024 Update: Pollen addition and exclusion

Reproduction in plants can be limited by access to pollen and resources. We previously found that Echinacea plants in the remnants are pollen limited, meaning that if they had access to more pollen, they would produce more seeds. However, the long-term effects of pollen limitation are unknown. Do plants that are super pollen saturated and have high amounts of pollen have a higher lifetime fitness than plants that are pollen limited? Also, we know that the plants in the remnants are pollen limited, but are the plants in the common garden environment also pollen limited? To answer these questions and more, 13 years ago Gretel randomly selected 39 plants from p1; half of these plants were randomly assigned to the pollen addition group, and the others were assigned to pollen exclusion. Every year, plants in the pollen exclusion have their heads bagged and they are not pollinated, while we hand cross every style in the pollen addition group. An additional 53 plants in p1 were selected to be part of a control group where pollen was neither added nor excluded.

In the summer of 2024, NONE of the original 39 addition/exclusion plants were flowering. If any had been flowering, the exclusion treatment plants would be covered with exclusion bags to prevent pollination, and the addition plants would be hand-pollinated multiple times throughout the summer. One plant in the control group produced a single flowering head.

Collaborator and NDSU postdoc Grace Hirzel places pollinator exclusions bags on Echinacea heads in the remnants. If any of the plants in the polLim experiment from the non-control group had flowered this year, it would’ve looked like this!

You can find more information about the pollen addition and exclusion experiment and links to previous flog posts regarding this experiment at the background page for the experiment.

Live from EntSoc!

Hey all!

It has been a packed few days here in Phoenix, and I’d like to describe some of the presentations I saw at yesterday’s ground nesting bee symposium. There was a broad range of topics, including the microbe communities found on pollen balls, brood parasite and host interactions, rare desert bees, and more! Nick Dorian had great results to share from his mark-recapture work in New Jersey, which was great to see after he demonstrated that method to Team Echinacea over the summer. Lily Fulton’s presentation really stood out to me: She is also studying prescribed fire, at a long term experimental fire site in Florida longleaf pine flatwoods. they found higher bee and wasp abundances during burn years, as well as higher proportions of bare ground. This is strikingly similar to our research findings! It was great to chat with her about it over my own poster.

Title slide of Lily Fulton’s presentation.
My poster! It is showing the first year of data from the ENRTF project, since we haven’t sent our 2024 bees to Zach yet.

It’s been incredible getting to meet so many researchers who care about native bees. Many stopped by to have a look at the poster! I’ve cited many of them in my previous work, so running into them in person is almost like meeting a celebrity. One consistent interest I’ve gotten from attendees is to look a the community composition of bees in the different treatment groups. Perhaps the increased abundance in the year of a burn we’re seeing is because of a cohort of burn-loving ground nesters, who don’t stick around in following years. It’ll be exciting to look into!

Shot of the poster sessions. There were a ton of applications, so space was pretty limited.

Overall, this has been a fantastic experience and opportunity to expand my network. I got to reconnect with some old friends, and make plenty of new ones. I can’t wait to get back to work on my thesis, now that I know who might be viewing it once it’s published! Expect more on that in the next few months.

Pollen and Nectar Update!

Exciting things are happening with Coreopsis pollen and nectar! Data entry for nectar has been accomplished and a figure has been made! The primary goal of this project is to see if prescribed burns have an affect on pollen and nectar quantity in C. palmata. However first and foremost, I want to go into more detail of how I collected data this summer, and then I’ll talk about my new nectar figure and what our findings are currently looking like.

Field Methods Step by Step:
Pre-Collection:

  • In order to collect pollen and nectar samples we first need to identify the plants we are sampling from.
  • Random bb-points are pre-selected to designate areas of study interest within each site. Identify the closest “patch”, which is a central location with 5 or more stems of coreopsis, relevant to the bb-point. Record the location of each patch using Avenza. (2024 Avenza layer can be found in “Dropbox / teamEchinacea2024 / maddieSadler / coreopsisPalmata”)
  • Place a flag in the relative center of the patch and label it with it’s patch ID number.
  • Select 5 plants that are still completely immature and place a pollen excluder bag over the head of the plant.
  • Monitor plants for a few days until they reach day two or three of anthesis where anthers are protruding and there are still immature florets in the center of the head. This is important later for pollen collection.

Pollen Collection:

  • Select the plant to sample from bagged plants located in each patch. We chose 3 of the 5 bagged plants based on day of anthesis and general look of the head.
  • Record the bb-point of the coreopsis patch, site name, and location on the data sheet.
  • Remove the pollinator exclusion bag from the selected head.
  • For pollen collection we will be collecting 3 immature florets from the head of the flower.
  • Label your microfuge tube with the plant ID number located on the data sheet.
  • Take the tweezers and carefully extract three immature florets, one by one, from the flower head and place them into the microfuge tube. Make sure to be careful that you do not rip the floret in half or puncture it with the tweezers.
  • Once all three florets are placed inside the tube, close it. Place the tube into the cooler with ice packs for further sorting upon your return from the field.
  • Repeat steps 1-8 for next plant.
  • Upon returning to the Hjelm House, place the collected pollen tubes in the collected samples box, which then is stored in the freezer.

Nectar Collection: This was adapted from the 2022 nectar protocol for Echinacea which can be found here.

  • After pollen collection you will begin the process of nectar collection.
  • On the same heads used from pollen collection, select the anthers that are the most recently presented to sample from.
  • Insert the microcap tube into the anther floret. Insert the microcap down into the floret until there is light resistance when you reach the base of the floret. 
  • Twist or rotate the microcap five times. 
  • Carefully remove the microcap.
  • Repeat steps 3-4 on all presented anther florets on the selected flower head.
  • Record the amount of nectar collected in the microcap in millimeters (mm). It is helpful to hold the microcap up to the sun to see the refraction of light from the nectar to see the amount collected. You can use a magnifying glass if needed to read the amount of nectar in mm on the ruler.
  • Place the entire microcap with the collected nectar sample into a microfuge tube to be disposed of properly outside of the field. 
  • Once this task is completed, it will not need to be done for the same plant in the future.
  • Repeat steps 2-8 for the next plant in the patch.
  • After all plants in patch are sampled from remove any extra pollinator bags and remove the flag from the center of the patch. Collection will not be repeated on the patch.

Field Supply Checklist:

Pollen Supplies:

  • Microfuge tubes (tall enough to put the immature floret inside and close the cap)
  • Permanent marker
  • Pen
  • Extra pollinator exclusion bags
  • Flag bag with flags of the designated color 
  • Magnifying glass glasses with 3.5 – 5 X magnification
  • Tweezers
  • Water and sunscreen 
  • Clipboard with the data sheet
  • Field collection box to hold small supplies 
  • Small cooler with ice packs

Nectar Supplies:

  • Microfuge tubes (tall enough to put the microcap inside and close the cap)
  • Microcaps: Drummond Scientific Microcap 1-000-0005 Microliter Pipets, 0.5 µl Capacity, 0.0056 inch diameter

These guys right here!

  • Extra pollinator exclusion bags
  • Flag bag with flags of the designated color 
  • Permanent marker
  • Pen
  • Magnifying glass glasses with 3.5 – 5 X magnification
  • Ruler with mm markings 
  • Water and sunscreen 
  • Clipboard with the data sheet
  • Field collection box to hold small supplies 

Now for the fun stuff!

Over this past week I’ve worked on creating this graph seen down below. This graph looks at the total amount of nectar in millimeters in each tube from each plant in our burned and unburned site combinations. These site combinations were created based on proximity to each other and burn history. For example, TorgN was burned, but TorgS, directly across from it, was not; Tower was burned, but Nice, directly across from it, was not. For YOHW and YOHE, we ran into an issue in that YOHE, the unburned side, had no flowering C. palmata in it; thus, we only have data from YOHW.

As we can see there is large amount of variation in totals across all the sites. When looking at the mean values (the red and blue squares on the graph) we are finding the slightest bit of evidence that burned sites are showing higher levels of nectar. Meaning my original hypothesis, that we’d see strong evidence that there is difference in quantity in burned sites rather than unburned sites is out the window! However, we can’t fully accept the null hypothesis, that there is strong evidence of no difference in nectar quantity, since there is not enough supporting evidence. Having this knowledge now, it will be interesting to see if there is a similar pattern in our pollen counts. Data and analysis on that to come!

Fig: Total (mm) of nectar in tubes for each plant at each site. Red squares (burned) and Blue squares (unburned) show average total with standard error.

**The code for this graph can be found in: “Dropbox / teamEchinacea2024 / maddieSadler / coreopsisPalmata”.**

NOTE: Funding for this project was provided by the Minnesota Environment and Natural Resources Trust Fund as recommended by the Legislative-Citizen Commission on Minnesota Resources (LCCMR).

rem 2024 Ea harvest summary

In 2024, we harvested Echinacea in the remnants for Jennifer Ison’s pollinator observation study and at the Hegg Lake pallida restoration.

For the pollinator observation study, we quantified reproductive effort on a given day and watched heads at different plants across 10 remnants. When a pollinator visited our focus head, we would take a video so we could later ID the pollinator.

This pollinator observation was done by Ning on a plant at yohw this summer. I asked Ian to give his best ID and he thinks this is a Halictus species! Very cool.

We harvested heads from plants involved in this experiment to get an idea of how observed visitation links up with realized seed set. In total, we harvested 136 heads and brought them back to the botanic garden for processing. Here are the detailed stats:

  • TOTAL: 136 heads
  • hulze: 15 heads (11.0%)
  • hulzw: 14 heads (10.3%)
  • hutche: 9 heads (6.6%)
  • hutchw: 13 heads (9.6%)
  • nice: 15 heads (11.0%)
  • torgen: 14 heads (10.3%)
  • torgew: 15 heads (11.0%)
  • tower: 11 heads (8.1%)
  • yohe: 15 heads (11.0%)
  • yohw: 15 heads (11.0%)

Again, please forgive rounding errors. This batch is the highest priority in the lab; volunteers are currently working on cleaning these heads and hopefully the rest of the ace process is soon to follow!

We also harvested, or perhaps I should just say decapitated, Echinacea pallida at Hegg Lake WMA. This species is native to areas primarily south and east of our study site, such as Indiana and Arkansas. SO how did they wind up in Minnesota? Seed for pallida was accidentally included in a restoration effort by the MN DNR years back. This raised concerns for us about potential hybridization with our native Echinacea angustifolia. (If you want to learn more about angustifolia x pallida hybridization like we did, check out our experiments to that end!) In summer 2024, we once again conducted our civic duty and decapitated pallida heads at Hegg in an attempt to prevent reproduction and further spread. We sliced and diced a total of 523 heads this year, with the largest plant having 20 normal flowering heads! Wowza.

Team members Liam and Maddie search for and take demography records of Echinacea pallida at Hegg lake before decapitating any heads that could later produce pollen. Photo credit Zach Zarling

There was also one plant at the location we refer to as “near pal” that we suspected may be an escaped hybrid; that is, a plant that we believe may be a natural hybrid of angustifolia and pallida. This individual only had one head, but was taller and more robust than its angustifolia neighbors. We put a pollinator exclusion bag on the head to prevent pollen spread, and later harvested the head and brought it back to the lab, where it is currently in the seed dryer. Keep an eye on 29239 in the future…